Principle of the Assay: This assay is a sandwich Enzyme Linked-Immunosorbent Assay (ELISA) for the measurement of human Nampt in biological fluids. A monoclonal antibody specific for Nampt has been precoated onto the 96-well microtiter plate. Standards and samples are pipetted into the wells for binding to the coated antibody. After extensive washing to remove unbound compounds, Nampt is recognized by the addition of a purified polyclonal antibody specific for Nampt (Detection Antibody). After removal of excess polyclonal antibody, HRP conjugated anti-rabbit IgG (HRP) is added. Following a final washing, peroxidase activity is quantified using the substrate 3,3',5,5'-tetramethylbenzidine (TMB). The intensity of the color reaction is measured at 450 nm after acidification and is directly proportional to the concentration of Nampt in the samples!!Background/Introduction: Fukuhara et al. (1) isolated visfatin, an adipocytokine that is highly enriched in the visceral fat of both humans and mice and whose expression level in plasma increases during the development of obesity. Visfatin corresponds to pre-B cell colony-enhancing factor (PBEF), a 52-kD cytokine expressed in lymphocytes. The gene encoding PBEF was originally isolated from an activated lymphocyte cDNA library (2). Although PBEF lacks a typical signal sequence for secretion, transfected COS-7 and mouse embryonic fibroblasts secreted PBEF into the culture medium. Samal et al. (2) found that recombinant PBEF secreted from transfected COS-7 and mouse embryonic fibroblasts was not itself active in a pre-B-cell colony formation assay, but it synergized the pre-Bcell colony formation activity of stem cell factor and interleukin-7. Jia et al. (3) found that PBEF is an inflammatory cytokine that plays a requisite role in the delayed neutrophil apoptosis of sepsis. Visfatin exerted insulin-mimetic effects in cultured cells and lowered plasma glucose levels in mice. Mice heterozygous for a targeted mutation in the visfatin gene had modestly higher levels of plasma glucose relative to wild type littermates. Surprisingly, it was found that visfatin binds to and activates the insulin receptor (1). However, this original discovery has not been reproduced by two groups (4-5). Visfatin, which is a secretory form of Nampt (nicotinamide phosphoribosyl-transferase), the ratelimiting enzyme of the mammalian NAD, plays a key role in secretion of insulin in the pancreatic beta-cells (5). Recently, two recent studies showed that plasma or serum levels of visfatin in individuals with type 2 diabetes mellitus was elevated (6-7), suggesting that measurement of plasma visfatin provides a relevant tool for understanding metabolic diseases.
NCBI and Uniprot Product Information
NCBI Description
This gene encodes a protein that catalyzes the condensation of nicotinamide with 5-phosphoribosyl-1-pyrophosphate to yield nicotinamide mononucleotide, one step in the biosynthesis of nicotinamide adenine dinucleotide. The protein belongs to the nicotinic acid phosphoribosyltransferase (NAPRTase) family and is thought to be involved in many important biological processes, including metabolism, stress response and aging. This gene has a pseudogene on chromosome 10. [provided by RefSeq, Feb 2011]
Uniprot Description
PBEF: an ubiquitous protein originally described as a cytokine which acts on early B-lineage precursor cells. Other reports indicate that PBEF is not a cytokine-like secreted protein but an intracellular protein associated with the cell cycle. Contains a catalytically active nicotinate phosphoribosyltransferase domain, an enzyme involved in nicotinamide adenine dinucleotide (NAD) biosynthesis.
Protein type: Cofactor and Vitamin Metabolism - nicotinate and nicotinamide; Cell cycle regulation; Transferase; Cytokine; EC 2.4.2.12
Chromosomal Location of Human Ortholog: 7q22.3
Cellular Component: cell junction; cytosol; extracellular space; nucleoplasm
Molecular Function: cytokine activity; drug binding; nicotinamide phosphoribosyltransferase activity; nicotinate-nucleotide diphosphorylase (carboxylating) activity; protein binding; protein homodimerization activity
Biological Process: cell-cell signaling; circadian regulation of gene expression; circadian rhythm; female pregnancy; insulin receptor signaling pathway; NAD biosynthetic process; NAD metabolic process; nicotinamide metabolic process; positive regulation of cell proliferation; positive regulation of nitric-oxide synthase biosynthetic process; positive regulation of smooth muscle cell proliferation; positive regulation of transcription from RNA polymerase II promoter; response to organic cyclic substance; signal transduction; vitamin metabolic process; water-soluble vitamin metabolic process
Research Articles on Nampt
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Product Notes
The Nampt nampt (Catalog #AAA495340) is an ELISA Kit and is intended for research purposes only. The product is available for immediate purchase. It is sometimes possible for the material contained within the vial of "Nampt, ELISA Kit" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.Precautions
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